Agent having neurotrophic factor-like activity

ABSTRACT

The present invention provides a pharmaceutical agent having high safety and a neurotrophic factor-like activity, which contains, as an active ingredient, any one compound included in fatty acids each having 8 carbon atoms (C8) or having 10 carbon atoms (C10) to 12 carbon atoms (C12) or fatty acid esters thereof, such as 3,7-dimethyloctanoic acid ethyl ester, geranic acid ethyl ester, and the like, each of which has 8 carbon atoms (C8), decanoic acid methyl ester, trans-2-decenoic acid, trans-2-decenoic acid methyl ester, trans-2-decenoic acid ethyl ester, trans-2-decenoic acid-2-decenyl ester, trans-2-decenoic acid cyclohexyl ester, trans-2-decenoic acid isopropyl ester, trans-3-decenoic acid methyl ester, trans-9-decenoic acid methyl ester, and the like, each of which has 10 carbon atoms (C10), trans-10-undecenoic acid methyl ester, trans-10-undecenoic acid ethyl ester, and the like, each of which has 11 carbon atoms (C11), and dodecanoic acid, and the like, each of which has 12 carbon atoms (C12), or salts thereof or prodrugs thereof.

TECHNICAL FIELD

The present invention relates to an agent having a neurotrophicfactor-like activity, which is useful for nervous disorders such asneurodegenerative diseases and depression due to promotion of activationof nerve cells, by activating signal transmission with a neurotrophicfactor-like activity of a nervous growth factor (NGF), a brain-derivedneurotrophic factor (BDNF), and the like.

BACKGROUND ART

A nerve cell is a cell having an information transmission function, andits damage emerges as serious loss of a cranial nerve function.Regeneration of an axon can hardly be expected in central nerves of abrain and spinal cord, and thus, when nerve cells have damage ordegeneration, activation of the nerve cells is required. A neurotrophicfactor is indispensable for activation of nerve cells of central nervesand peripheral nerves such as differentiation of nerve cells, survivalsustention, promotion of synapse functions, regeneration and restorationin damage of nerve cells, and the like. Among neurotrophic factors, anervous growth factor (NGF), a brain-derived neurotrophic factor (BDNF),neurotrophin-3 (NT-3), neurotrophin-4/5 (NT-4/5), and the like constructa neurotrophin family having 50% or more of sequence homology with anervous growth factor (NGF) as a prototype. Activation of nerve cells isachieved by bonding a neurotrophic factor secreted outside the cellswith a high-affinity receptor (Trks) to activate signal transmissionthrough a MAP kinase (mitogen-activated protein (MAP) kinase)information transmission path that activates (phosphorylates) MAPkinases in the nerve cells, and controlling numerous genetic expressionsthrough activation of CREB (c-AMP-response element binding protein) of atranscription factor, and the like.

Therefore, if signal transmission through a MAP kinase informationtransmission path can be activated, there is a possibility of clinicalapplications to a nervous disorder caused by degeneration of nerve cellsor cellular death. Further, there is a report about a relation betweenbrain-derived neurotrophic factors (BDNF) and some diseases. On thebasis of studies using a polymorphism of a brain-derived neurotrophicfactor (BDNF), there are a report that a brain-derived neurotrophicfactor (BDNF) is associated with Parkinson's disease (see Non-patentDocument 1), a report that a brain-derived neurotrophic factor (BDNF) isassociated with Alzheimer's disease (see Non-patent Document 2), areport that a brain-derived neurotrophic factor (BDNF) is associatedwith depression (see Non-patent Document 3), a report that abrain-derived neurotrophic factor (BDNF) is associated with bipolardepression (see Non-patent Document 4), and a report that abrain-derived neurotrophic factor (BDNF) is associated with anxietydisorder (see Non-patent Document 5). Furthermore, there are a reportthat decrease in a synapse function of a genetically converted mousehaving Huntington's disease is cured with administration of abrain-derived neurotrophic factor (BDNF) (see Non-patent Document 6),and a report that administration of a MAP kinase phosphorylationinhibitor provokes an antidepressant condition (see Non-patent Document7). Accordingly, neurotrophic factors are expected to have an effect asa therapeutic agent for nervous disorders. However, since neurotrophicfactors are polymer proteins, they have a problem of having difficultyin reaching the brain since they cannot pass through a blood-brainbarrier even if administered from a peripheral. Thus, it has been triedto search medical drugs having a neurotrophic factor-like activity thatactivates nerve cells with a low-molecular weight compound and medicaldrugs promoting production and secretion of neurotrophic factors.

Conventionally, an agent having a neurotrophic factor-like activitycontaining a compound having a predetermined general formula has beenproposed (Patent Documents 1 and 2). A production and secretionaccelerator of a neurotrophic factor containing a compound having apredetermined general formula (see Patent Documents 3 to 5) and a nerveregeneration accelerator containing fatty acid compounds, salts thereofor prodrugs thereof (see Patent Document 6) have been proposed.

Furthermore, there has been a proposal for a pharmaceutical agentcontaining a compound having a predetermined general formula, whichprevents and cures neurodegenerative diseases with improvement ofdecrease in GABAA receptor response of an astrocyte (see Patent Document7).

-   Non-patent Document 1: Ann Neurol. January 2002; 51(1): 133-6-   Non-patent Document 2: J Neural Transm. May 2005; 112(5) : 703-11.    Epub Sep. 14, 2004-   Non-patent Document 3: Neuropsychopharmacology. February 2003;    28(2): 397-401. Epub Aug. 29, 2002-   Non-patent Document 4: Br J Psychiatry. October 2006; 189: 317-23-   Non-patent Document 5: Psychopharmacology (Berl). June 2005; 180(1):    95-9. Epub Jan. 26, 2005-   Non-patent Document 6: J Neurosci. Apr. 18, 2007; 27(16): 4424-34-   Non-patent Document 7: BIOL PSYCHIATRY 2007; 61: 661-670-   Patent Document 1: Japanese Unexamined Patent Application (JP-A) No.    2000-7568-   Patent Document 2: JP-A No. 2003-113085-   Patent Document 3: JP-A No. 2002-80467-   Patent Document 4: JP-A No. 2003-261545-   Patent Document 5: WO No. 2003/084542-   Patent Document 6: WO No. 2005/032535-   Patent Document 7: JP-A No. H07-316092

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

The agents having neurotrophic factor-like activities or the productionand secretion accelerators of neurotrophic factors described in PatentDocuments 1 to 5 do not contain fatty acids or fatty acid esters asactive ingredients. In the nerve regeneration accelerator described inPatent Document 6, an active ingredient disclosed with a pharmacologicalactivity of nerve regeneration is (2R)-2-propyloctanoic acid. Thepharmaceutical agent for preventing and curing neurodegenerativediseases described in Patent Document 7 contains a saturated fatty acidhaving 10 or less carbon atoms (C10), an unsaturated fatty acid orsaturated fatty acid ester each having 5 carbon atoms (C5) as an activeingredient.

An object of the present invention is to provide a pharmaceutical agenthaving high safety and a neurotrophic factor-like activity, whichcontains a predetermined fatty acid or fatty acid ester as an activeingredient.

Means for Solving the Problems

A gist of the present invention lies in an agent having a neurotrophicfactor-like activity, containing, as an active ingredient, anyonecompound included in fatty acids each having 8 carbon atoms (C8) orhaving 10 carbon atoms (C10) to 12 carbon atoms (C12) or fatty acidesters thereof, such as n-octanoic acid methyl ester, n-octanoic acidethyl ester, 3,7-dimethyloctanoic acid ethyl ester, or geranic acidethyl ester, each of which has 8 carbon atoms (C8), decanoic acid methylester, decanoic acid ethyl ester, trans-2-decenoic acid,trans-2-decenoic acid methyl ester, trans-2-decenoic acid ethyl ester,trans-2-decenoic acid-2-decenyl ester, trans-2-decenoic acid cyclohexylester, trans-2-decenoic acid octyl ester, trans-2-decenoic acidisopropyl ester, trans-3-decenoic acid methyl ester, trans-3-decenoicacid ethyl ester, trans-9-decenoic acid, rans-9-decenoic acid methylester, or trans-9-decenoic acid ethyl ester, each of which has 10 carbonatoms (C10), trans-10-undecenoic acid methyl ester, ortrans-10-undecenoic acid ethyl ester, each of which has 11 carbon atoms(C11), and dodecanoic acid, dodecanoic acid methyl ester, or dodecanoicacid ethyl ester, each of which has 12 carbon atoms (C12), or saltsthereof or prodrugs thereof.

A gist of the present invention lies in the agent having a neurotrophicfactor-like activity for a preventive and therapeutic agent for anervous disorder. The nervous disorder may be a neurodegenerativedisease such as Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis (ALS), Huntington's disease, progressive supranuclearpalsy (PSP), or diabetic neuropathy. The nervous disorder may be amental disease, and the mental disease may be depression or anxietydisorder (neurosis).

A gist of the present invention lies in trans-2-decenoic acid-2-decenylester, having a neurotrophic factor-like activity. A gist of the presentinvention lies in trans-2-decenoic acid cyclohexyl ester, having aneurotrophic factor-like activity.

Effect of the Invention

The agent having a neurotrophic factor-like activity of the presentinvention is useful as a preventive and therapeutic agent for a nervousdisorder, which has high safety and activates signal transmissionthrough a MAP kinase information transmission path. The agent is usefulas a preventive and improving agent for a neurodegenerative disease suchas Alzheimer's disease, Parkinson's disease, amyotrophic lateralsclerosis (ALS), Huntington's disease, progressive supranuclear palsy(PSP), or diabetic neuropathy in nervous disorders. The agent is usefulas a preventive and improving agent for a mental disease in the nervousdisorder. The agent is useful as a preventive and improving agent fordepression or anxiety disorder (neurosis) in the mental disease. Inparticular, the agent can be expected to have an immediateantidepressant effect and antianxiety effect as a preventive andimproving agent for depression or anxiety disorder (neurosis).

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a graph showing ratios of phosphorylated MAP kinases to MAPkinases of compounds 1, 2, 3, 6, 7, 8, 11, 12 and 13 in Example 1.

FIG. 2 is a graph showing ratios of phosphorylated MAP kinases to MAPkinases of compounds 16, 17, 18, 21, 22, 23, 36, 37 and 38 in Example 1.

FIG. 3 is a graph showing ratios of phosphorylated MAP kinases to MAPkinases of compounds 47, 48, 53, 54, 62, 63, 77 and 78 in Example 1.

FIG. 4 is a graph of a burying the glass-marbles test of comparing thenumbers of buried glass marbles between stressed examples andnon-stressed examples of the “stressed 3 week-administration groups” inExample 2.

FIG. 5 is a graph of a burying the glass-marbles test of comparing thenumbers of burying glass marbles between stressed examples andnon-stressed examples of the “stressed 1 week-administration groups” inExample 2.

FIG. 6 is a graph of a tail suspension test of comparing immobilitytimes between stressed examples and non-stressed examples of the“stressed 3 week-administration groups” in Example 2.

FIG. 7 is a graph of a tail suspension test of comparing immobilitytimes between stressed examples and non-stressed examples of the“stressed 1 week-administration groups” in Example 2.

FIG. 8 is a graph of an elevated plus maze test of comparing times forbeing in open arms between stressed examples and non-stressed examplesof the “stressed 3 week-administration groups” in Example 2.

FIG. 9 is a graph of an elevated plus maze test of comparing the numbersof times of entering closed arms between stressed examples andnon-stressed examples of the “stressed 3 week-administration groups” inExample 2.

FIG. 10 is a graph of an elevated plus maze test of comparing times forbeing in open arms between stressed examples and non-stressed examplesof the “stressed 1 week-administration groups” in Example 2.

FIG. 11 is a graph of an elevated plus maze test of comparing thenumbers of times of entering closed arms between stressed examples andnon-stressed examples of the “stressed 1 week-administration groups” inExample 2.

FIG. 12 is a graph comparing gaps of body weights between stressedexamples obtained by administering the compound 8 and applying stressfor 3 weeks and non-stressed examples in Example 3.

FIG. 13 is a graph comparing phosphorylated MAP kinase amounts incortexes of frontal lobes between stressed examples obtained byadministering the compound 8 and applying stress for 3 weeks andnon-stressed examples in Example 4.

FIG. 14 is a graph comparing phosphorylated MAP kinase amounts inhippocampi between stressed examples obtained by administering thecompound 8 and applying stress for 3 weeks and non-stressed examples inExample 4.

FIG. 15 is a graph comparing expression amounts of mRNAs of BDNF, NT-3,NGF and GR in cortexes of the frontal lobes between stressed examplesobtained by administering the compound 8 and applying stress for 3 weeksand non-stressed examples in Example 5.

FIG. 16 is a graph comparing expression amounts of mRNAs of BDNF, NT-3,NGF and GR in hippocampi between stressed examples obtained byadministering the compound 8 and applying stress for 3 weeks andnon-stressed examples in Example 5.

FIG. 17 is a graph of a burying the glass-marbles test of comparing thenumbers of buried glass marbles between stressed examples andnon-stressed examples of the “non-administration groups after completionof stress” in Example 6.

FIG. 18 is a graph of a burying the glass-marbles test of comparing thenumbers of buried glass marbles between stressed examples andnon-stressed examples of the “1 week-administration groups aftercompletion of stress” in Example 6.

FIG. 19 is a graph of a burying the glass-marbles test of comparing thenumbers of buried glass marbles between stressed examples andnon-stressed examples of the “2 week-administration groups aftercompletion of stress” in Example 6.

FIG. 20 is a graph of a tail suspension test of comparing immobilitytimes between stressed examples and non-stressed examples of the“non-administration groups after completion of stress” in Example 6.

FIG. 21 is a graph of a tail suspension test of comparing immobilitytimes between stressed examples and non-stressed examples of the “1week-administration groups after completion of stress” in Example 6.

FIG. 22 is a graph of a tail suspension test of comparing immobilitytimes between stressed examples and non-stressed examples of the “2week-administration groups after completion of stress” in Example 6.

FIG. 23 is a graph of an elevated plus maze test of comparing times forbeing in open arms between stressed examples and non-stressed examplesof the “non-administration groups after completion of stress” in Example6.

FIG. 24 is a graph of an elevated plus maze test of comparing thenumbers of times of entering closed arms between stressed examples andnon-stressed examples of the “non-administration groups after completionof stress” in Example 6.

FIG. 25 is a graph of an elevated plus maze test of comparing times forbeing in open arms between stressed examples and non-stressed examplesof the “1 week-administration groups after completion of stress” inExample 6.

FIG. 26 is a graph of an elevated plus maze test of comparing thenumbers of times of entering closed arms between stressed examples andnon-stressed examples of the “1 week-administration groups aftercompletion of stress” in Example 6.

FIG. 27 is a graph of an elevated plus maze test of comparing times forbeing in open arms between stressed examples and non-stressed examplesof the “2 week-administration groups after completion of stress” inExample 6.

FIG. 28 is a graph of an elevated plus maze test of comparing thenumbers of times of entering closed arms between stressed examples andnon-stressed examples of the “2 week-administration groups aftercompletion of stress” in Example 6.

FIG. 29 is a graph comparing gaps of body weights between stressedexamples immediately after applying stress for 3 weeks and non-stressedexamples in Example 7.

FIG. 30 is a graph comparing gaps of body weights between stressedexamples obtained by administering the compound 8 for 1 week afterapplying stress for 3 weeks and non-stressed examples in Example 7.

FIG. 31 show an evaluation method of a behavior test (behaviorassessment) (A) and a graph of a tail suspension test (B) comparingeffects on antidepressant symptoms with the compound 8 and fluvoxaminein Example 8.

FIG. 32 are graphs of a voluntary alternation behavior test (Y-mazetest) of mice administered with trimethyltin in Example 9. FIG. 32A is agraph showing ratios of alternation behavior (%) of respective testgroups, and FIG. 32B is a graph showing the numbers of total armselection (total arm entries) of respective test groups.

FIG. 33 are graphs of a novel object recognition test of mice in Example10. FIG. 33A is a graph showing time for the mice to explore a “familiarobject” and a “novel object” (exploration time), and FIG. 33B is a graphshowing recognition indices.

BEST MODE FOR CARRYING OUT THE INVENTION

Commercially available products can be used as a fatty acid that is anactive ingredient of the agent having a neurotrophic factor-likeactivity of the present invention, or can be produced in known methodsin the same manner as the commercially available products. A fatty acidester that is an active ingredient of the agent having a neurotrophicfactor-like activity of the present invention can be produced in knownmethods. For examples, it can be produced by the Fischer esterificationmethod in which a fatty acid and an alcohol are reacted in the presenceof an acid catalyst, or a transesterification method. Furthermore, itcan be produced by substituting a hydroxyl group of a fatty acid to ahalogen with a thionyl chloride, sulfiryl chloride, phosphorustrichloride, phosphorus pentachloride, oxalyl chloride, phosphorictrichloride, or the like to give acyl halide and reacting the acylhalide with an alcohol. The produced fatty acid ester can be isolatedusing known separation and purification techniques such as extraction,partition, and column chromatography, after completion of the reaction.

The fatty acid or the fatty acid ester is any one compound included infatty acids each having 8 carbon atoms (C8) or having 10 carbon atoms(C10) to 12 carbon atoms (C12) or fatty acid esters thereof, such asn-octanoic acid methyl ester, n-octanoic acid ethyl ester,3,7-dimethyloctanoic acid ethyl ester, or geranic acid ethyl ester, eachof which has 8 carbon atoms (C8), decanoic acid methyl ester, decanoicacid ethyl ester, trans-2-decenoic acid, trans-2-decenoic acid methylester, trans-2-decenoic acid ethyl ester, trans-2-decenoicacid-2-decenyl ester, trans-2-decenoic acid cyclohexyl ester,trans-2-decenoic acid octyl ester, trans-2-decenoic acid isopropylester, trans-3-decenoic acid methyl ester, trans-3-decenoic acid ethylester, trans-9-decenoic acid, rans-9-decenoic acid methyl ester, ortrans-9-decenoic acid ethyl ester, each of which has 10 carbon atoms(C10), trans-10-undecenoic acid methyl ester, or trans-10-undecenoicacid ethyl ester, each of which has 11 carbon atoms (C11), anddodecanoic acid, dodecanoic acid methyl ester, or dodecanoic acid ethylester, each of which has 12 carbon atoms (C12). One kind or combinationof two kinds or more of these compounds can be suitably used.

The fatty acids or fatty acid esters can also be isolated fatty acids orfatty acid esters, or pharmaceutically acceptable salts thereof orprodrugs thereof. Examples of the salts include alkali metal salts suchas sodium salt and potassium salt; alkali earth metal salts such asmagnesium salt and calcium salt; inorganic acids such as hydrochloricacid, sulfuric acid, phosphoric acid, and hydrobromic acid; alkanolamines such as ammonium salt and triethanol amine; and acid additionsalts of organic acids such as acetic acid, formic acid, fumaric acid,and oxalic acid. The fatty acids or fatty acid esters can be solvates ofthe fatty acids or fatty acid esters. Examples of the solvates includehydrates and alcoholates.

The agent having a neurotrophic factor-like activity is useful forprevention and treatment of nervous disorders. The nervous disorderrefers to a clinical condition of damaging functions of nerve cells dueto degeneration and cell death of the nerve cells, and includesneurodegenerative diseases and mental diseases. The neurodegenerativediseases refer to Alzheimer's disease, Parkinson's disease, amyotrophiclateral sclerosis (ALS), Huntington's disease, progressive supranuclearpalsy (PSP), diabetic neuropathy, etc. The mental diseases refer todepression (including bipolar depression), anxiety disorder (neurosis),integration disorder syndrome, etc. When the agent is used fordepression, it takes at least 3 to 4 weeks until there appear effects ofconventionally existing depression therapeutic agents such as atricyclic antidepressant, a tetracyclic antidepressant, a selectiveserotonin reuptake inhibitor (SSRI), a serotonin noradrenalin reuptakeinhibitor (SNRI), and the like, and these agents had to be takenperiodically during this term; however, the agent having a neurotrophicfactor-like activity of the present invention can be expected to haveimmediate affectivity as compared to existing medical drugs.

An administration form of the agent having a neurotrophic factor-likeactivity as a medical drug is not particularly limited, and may be anyadministration form of oral and parenteral routes. A suitable dosageform can be employed according to an administration form, and the agentcan be formed into various preparations such as injectable agents, ororal agents (e.g., capsules, tablets, granules, sprays, pills, subtlegranules, etc.), rectal administration agents, oleagenous suppositories,and aqueous suppositories.

Various preparations are prepared by adding vehicles, binding agents,brighteners, disintegrating agents, surfactants, fluidity accelerators,and the like, which are pharmaceutically acceptable and generally used.Examples of the vehicles include lactose, fructose, glucose, cornstarch,sorbit, crystalline cellulose. Examples of the binding agents includemethyl cellulose, ethyl cellulose, gum arabic, gelatin, hydroxypropylcellulose, and polyvinyl pyrrolidone. Examples of the brightenersinclude talc, magnesium stearate, polyethylene glycol, and curingvegetable oils. Examples of the disintegrating agents include starch,sodium alginate, gelatin, calcium carbonate, calcium citrate, dextrin,magnesium carbonate, and synthetic magnesium silicate. Examples of thesurfactants include sodium lauryl sulfate, soybean lecithin, sucrosefatty acid esters, and polysorbate 80. Examples of the fluidityaccelerators include light anhydrous silicic acid, a dry aluminumhydroxide gel, synthetic aluminum silicate, and magnesium silicate.Examples of other additives include a syrup, vaseline, glycerin,ethanol, propylene glycol, citric acid, sodium chloride, sodium nitrite,and sodium phosphate.

An administration dose of the agent having a neurotrophic factor-likeactivity can be suitably increased in consideration of a usage, an age,a sex and a degree of a symptom of a patient, and the like, and isgenerally 1 to 1000 mg, and preferably 5 to 300 mg, per day for anadult, and such a dose can be administered once a day or dividedlyadministered several times a day.

Examples

The present invention will be then described in reference to examples,and the present invention is not limited to the following examples.

Production Example 1 Decanoic acid methyl ester, Compound 2

Decanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Methanol(30 ml) was added to chloride of decanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with ethyl acetate (hereinafterexpressed as “EtOAc”). The distributed liquid was purified with columnchromatography (developing solvent: n-hexane (C₆H₁₄)-EtOAc (3:1)) afterconcentration, and decanoic acid methyl ester shown in the following wasisolated.

Colorless oily substance, C₁₁H₂₂O₂ MW 186, EIMS m/z (%): 186 (M+, 7),143 (21), 101 (10), 87 (55), 74 (100)

Production Example 2 Decanoic acid ethyl ester, Compound 3

Decanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Ethanol(30 ml) was added to chloride of decanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with EtOAc. The distributedliquid was purified with column chromatography (developing solvent:C₆H₁₄-EtOAc (3:1)) after concentration, and decanoic acid ethyl estershown in the following was isolated.

Colorless oily substance, C₁₂H₂₄O₂ MW 200, EIMS m/z (%): 200 (M+, 10),155 (26), 101 (44), 88 (100), 73 (19)

Production Example 3 Trans-2-decenoic acid methyl ester, Compound 7

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Methanol (30 ml) was added to chloride of trans-2-decenic acidand refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-2-decenoic acid methyl ester shown in the following was isolated.

Colorless oily substance, C₁₁H₂₀O₂ MW 184, EIMS m/z (%): 184 (M+, 5),167 (6), 153 (43), 123 (16), 113 (41), 87 (90), 69 (39), 43 (100)

Production Example 4 Trans-2-decenoic acid ethyl ester, Compound 8

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Ethanol (30 ml) was added to chloride of trans-2-decenic acidand refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-2-decenoic acid ethyl ester shown in the following was isolated.

Colorless oily substance, C₁₂H₂₂O₂ MW 198, EIMS m/z (%): 198 (M+, 9),171 (27), 153 (56), 110 (11), 91 (100), 71 (44)

Production Example 5 Trans-9-decenoic acid methyl ester, Compound 12

Trans-9-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Methanol (30 ml) was added to chloride of trans-9-decenic acidand refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-9-decenoic acid methyl ester shown in the following was isolated.

Colorless oily substance, C₁₁H₂₀O₂ MW 184, EIMS m/z (%): 184 (M+, 0),166 (M+ —CH3OH, 37), 135 (28), 110 (38), 91 (80), 74 (100)

Production Example 6 Trans-9-decenoic acid ethyl ester, Compound 13

Trans-9-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Ethanol (30 ml) was added to chloride of trans-9-decenic acidand refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-9-decenoic acid ethyl ester shown in the following was isolated.

Colorless oily substance, C₁₂H₂₂O₂ MW 198, EIMS m/z (%): 198 (M+, 0),152 (M+ —C2H5OH, 13), 135 (17), 101 (17), 91 (100)

Production Example 7 Dodecanoic acid methyl ester, Compound 17

Dodecanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Methanol(30 ml) was added to chloride of dodecanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with EtOAc. The distributedliquid was purified with column chromatography (developing solvent:C₆H₁₄-EtOAc (3:1)) after concentration, and dodecanoic acid methyl estershown in the following was isolated.

Colorless oily substance, C₁₃H₂₆O₂ MW 214, EIMS m/z (%): 214 (M+, 13),185 (15), 171 (15), 143 (17), 87 (64), 74 (100)

Production Example 8 Dodecanoic acid ethyl ester, Compound 18

Dodecanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Ethanol(30 ml) was added to chloride of dodecanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with EtOAc. The distributedliquid was purified with column chromatography (developing solvent:C₆H₁₄-EtOAc (3:1)) after concentration, and dodecanoic acid ethyl estershown in the following was isolated.

Colorless oily substance, C₁₄H₂₈O₂ MW 228, EIMS m/z (%): 228 (M+, 17),183 (28), 157 (15), 101 (50), 88 (100), 7 (64), 74 (100)

Production Example 9 Trans-10-undecenoic acid methyl ester, Compound 22

Trans-10-undecenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Methanol (30 ml) was added to chloride of trans-10-undecenicacid and refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-10-undecenoic acid methyl ester shown in the following wasisolated.

Colorless oily substance, C₁₂ H₂₂O₂ MW 198, EIMS m/z (%): 198 (M+, 0),166 (M+ —CH3OH, 22), 149 (13), 124 (36), 87 (53), 74 (100)

Production Example 10 Trans-10-undecenoic acid ethyl ester, Compound 23

Trans-10-undecenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Ethanol (30 ml) was added to chloride of trans-10-undecenicacid and refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-10-undecenoic acid ethyl ester shown in the following wasisolated.

Colorless oily substance, C₁₃H₂₄O₂ MW 212, EIMS m/z (%): 212 (M+, 2),166 (M+ —CH3OH, 32), 149 (20), 124 (38), 101 (44), 88 (84), 69 (34), 41(100)

Production Example 11 n-octanoic acid methyl ester, Compound 37

n-octanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Methanol(30 ml) was added to chloride of n-octanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with EtOAc. The distributedliquid was purified with column chromatography (developing solvent:C₆H₁₄-EtOAc (3:1)) after concentration, and n-octanoic acid methyl estershown in the following was isolated.

Colorless oily substance, C₉H₁₈O₂ MW 158, EIMS m/z (%): 158 (M+, 4), 127(27), 115 (11), 88 (43), 74 (100)

Production Example 12 n-octanoic acid ethyl ester, Compound 38

n-octanoic acid was dissolved in thionyl chloride (10 ml) and treated inwater bath for 3 hours to distill off excess thionyl chloride. Ethanol(30 ml) was added to chloride of n-octanoic acid and refluxed in waterbath for 2 hours. After cooling, the reaction mixture was added to 1NHCl (80 ml) to be acidic and distributed with EtOAc. The distributedliquid was purified with column chromatography (developing solvent:C₆H₁₄-EtOAc (3:1)) after concentration, and n-octanoic acid ethyl estershown in the following was isolated.

Colorless oily substance, C₁₀H₂₀O₂ MW 172, EIMS m/z (%): 172 (M+, 6),127 (42), 115 (11), 101 (39), 88 (100)

Production Example 13 Trans-2-decenoic acid-trans-2-decenyl ester,Compound 47

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Trans-2-decen-1-ol (30 ml) was added to chloride oftrans-2-decenoic acid and refluxed in water bath for 2 hours. Aftercooling, the reaction mixture was added to 1N HCl (80 ml) to be acidicand distributed with EtOAc. The distributed liquid was purified withcolumn chromatography (developing solvent: C₆H₁₄-EtOAc (3:1)) afterconcentration, and trans-2-decenoic acid-trans-2-decenyl ester shown inthe following was isolated.

Colorless oily substance, C₂₀H₃₆O₂ MW 308, EIMS m/z (%): 308 (M+, 8),153 (100), 138 (12), 110 (10), 91 (36), 69 (24), 55 (35)

Production Example 14 Trans-2-decenoic acid cyclohexyl ester, Compound48

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Cyclohexanol (30 ml) was added to chloride of trans-2-decenoicacid and refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-2-decenoic acid cyclohexyl ester shown in the following wasisolated.

Colorless oily substance, C₁₆H₂₈O₂ MW 252, EIMS m/z (%): 252 (M+, 0),171 (100), 153 (30), 82 (16), 67 (10), 55 (13)

Production Example 15 Trans-2-decenoic acid octyl ester, Compound 53

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. 1-octanol (30 ml) was added to chloride of trans-2-decenoicacid and refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, andtrans-2-decenoic acid octyl ester shown in the following was isolated.

Colorless oily substance, C₁₈H₃₄O₂ MW 282, EIMS m/z (%): 282 (M+, 1.6),171 (100), 153 (18), 112 (14), 83 (25), 69 (22), 57(26)

Production Example 16 Trans-2-decenoic acid isopropyl ester, Compound 54

Trans-2-decenoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. An isopropyl alcohol (30 ml) was added to chloride oftrans-2-decenoic acid and refluxed in water bath for 2 hours. Aftercooling, the reaction mixture was added to 1N HCl (80 ml) to be acidicand distributed with EtOAc. The distributed liquid was purified withcolumn chromatography (developing solvent: C₆H₁₄-EtOAc (3:1)) afterconcentration, and trans-2-decenoic acid isopropyl ester shown in thefollowing was isolated.

Colorless oily substance, C₁₃H₂₄O₂ MW 212, EIMS m/z (%): 212 (M+, 1.6),171 (38), 153 (54), 110 (11), 99 (11), 57 (26), 43(100)

Production Example 17 Trans-3-decenoic acid methyl ester, Compound 62

Trans-3-decenoic acid (Wako Pure Chemical Industries, Ltd.) wasdissolved in thionyl chloride (10 ml) and treated in water bath for 3hours to distill off excess thionyl chloride. Methanol (30 ml) was addedto chloride of trans-3-decenoic acid and refluxed in water bath for 2hours. After cooling, the reaction mixture was added to 1N HCl (80 ml)to be acidic and distributed with EtOAc. The distributed liquid waspurified with column chromatography (developing solvent: C₆H₁₄-EtOAc(3:1)) after concentration, and trans-3-decenoic acid methyl ester shownin the following was isolated.

Colorless oily substance, C₁₁H₂₀O₂ MW 184, EIMS m/z (%): 183 (M+-1, 14),171 (30), 151 (8), 139 (25), 123 (46), 97 (25), 87 (62), 55 (47), 41(100)

Production Example 18 Trans-3-decenoic acid ethyl ester, Compound 63

Trans-3-decenoic acid (Wako Pure Chemical Industries, Ltd.) wasdissolved in thionyl chloride (10 ml) and treated in water bath for 3hours to distill off excess thionyl chloride. Ethanol (30 ml) was addedto chloride of trans-3-decenoic acid and refluxed in water bath for 2hours. After cooling, the reaction mixture was added to 1N HCl (80 ml)to be acidic and distributed with EtOAc. The distributed liquid waspurified with column chromatography (developing solvent: C₆H₁₄-EtOAc(3:1)) after concentration, and trans-3-decenoic acid ethyl ester shownin the following was isolated.

Colorless oily substance, C₁₂H₂₂O₂ MW 198, EIMS m/z (%): 197 (M+-1, 23),185 (31), 157(20), 139 (52), 123 (100), 101 (85), 73 (56)

Production Example 19 Geranic acid ethyl ester, Compound 77

Geranic acid (Wako Pure Chemical Industries, Ltd.) was dissolved inthionyl chloride (10 ml) and treated in water bath for 3 hours todistill off excess thionyl chloride. Ethanol (30 ml) was added tochloride of geranic acid and refluxed in water bath for 2 hours. Aftercooling, the reaction mixture was added to 1N HCl (80 ml) to be acidicand distributed with EtOAc. The distributed liquid was purified withcolumn chromatography (developing solvent: C₆H₁₄-EtOAc (3:1)) afterconcentration, and geranic acid ethyl ester shown in the following wasisolated.

Colorless oily substance, C₁₂H₂₀O₂ MW 196, EIMS m/z (%): 195 (M+-1, 17),181 (10), 151 (15), 121 (74), 107 (100), 79 (40), 69 (30)

Production Example 20 3, 7-dimethyloctanoic acid ethyl ester, Compound78

3,7-dimethyloctanoic acid was dissolved in thionyl chloride (10 ml) andtreated in water bath for 3 hours to distill off excess thionylchloride. Ethanol (30 ml) was added to chloride of 3,7-dimethyloctanoicacid and refluxed in water bath for 2 hours. After cooling, the reactionmixture was added to 1N HCl (80 ml) to be acidic and distributed withEtOAc. The distributed liquid was purified with column chromatography(developing solvent: C₆H₁₄-EtOAc (3:1)) after concentration, and the3,7-dimethyloctanoic acid ethyl ester shown in the following wasisolated. 3,7-dimethyloctanoic acid was obtained by contact reduction ofgeranic acid.

Colorless oily substance, C₁₂H₂₄O₂ MW 200, EIMS m/z (%): 200 (M+, 6),198 (10), 185 (6), 153 (22), 128 (24), 115 (68), 88 (100), 69 (45)

Example 1 Measurement of Activation of MAP Kinases

Activation of MAP kinases was measured on respective compounds in Table1 including the compound obtained in Production Example with westernimmunoblotting as follows. Nerve cells were dispersed from a cerebralcortex of a 17-day-old fetal rat and the nerve cells were cultured in aDulbecco's modified eagle medium (DMEM) containing 5% fetal bovine serumfor 1 day. A culture solution was exchanged to a serum-free medium (B27supplement added Neurobasal, Invitrogen Corporation), and the nervecells were cultured in a culture petri dish coated with polyornithine ata density of 20,000 to 40,000 cells/cm². Three days later, each of thecompounds in Table 1 was added thereto, and culture for 30 minutes wascontinued. Then, cells were recovered on ice with a solution of aTris-HCL buffer as a base containing a phosphatase inhibitor. Theprotein concentration of the obtained cell extraction was determinedusing a BCA Protein Assay Kit (TAKARABIO INC.), a constant amount (3 μgfor MAP kinase measurement, 5 μg for phosphorylated MAP kinasemeasurement) of the protein was electrophoresed with a polyacrylic amidegel. The protein was transcribed to a PVDF membrane from the gel afterelectrophoresis, and western immunoblotting was carried out using theprimary antibodies: an anti-MAP kinase antibody (Cell SignalingTechnology, Inc.) and an antiphosphorylated MAP kinase antibody (CellSignaling Technology, Inc.), respectively. Subsequently, an enzymeactivity was subjected to color development by being reacted with thesecondary antibody: an alkaline phosphatase labeled anti-rabbit IgGantibody (Promega KK) to measure MAP kinases and phosphorylated MAPkinases. In addition, each compound was dissolved in 0.1% DMSO andadjusted to have a concentration of 500 μg/ml. A control was added witha phosphoric acid buffer containing 0.1% DMSO in the same amount as thecompound.

The above obtained concentration measurement of the electrophoresis gelband was digitalized by calculating an intensity with Image J (BioArtsInternational, Inc.). The numerical value of MAP kinases of eachcompound was divided by the numerical value of the control's MAPkinases, and the numerical value of phosphorylated MAP kinases of eachcompound was divided by the numerical value of the control'sphosphorylated MAP kinases to find a ratio of the MAP kinases of eachcompound to the control and a ratio of the phosphorylated MAP kinases ofthe each compound to the control. Then, the obtained ratio of thephosphorylated MAP kinases to the control was divided by the obtainedratio of the MAP kinases to the control to find a ratio of thephosphorylated MAP kinases to the MAP kinases and graphs were formed andshown in FIGS. 1 to 3. Compound 1, compound 6, compound 11, compound 16,compound 21 and compound 36 in Table 1 were used from products made byWako Pure Chemical Industries, Ltd.

TABLE 1 Compound Nos. Compound 1 decanoic acid 2 decanoic acid methylester 3 decanoic acid ethyl ester 6 trans-2-decenoic acid 7trans-2-decenoic acid methyl ester 8 trans-2-decenoic acid ethlyl ester11 trans-9-decenoic acid 12 trans-9-decenoic acid methyl ester 13trans-9-decenoic acid ethyl ester 16 dodecanoic acid 17 dodecanoic acidmethyl ester 18 dodecanoic acid ethyl ester 21 trans-10-undecenoic acid22 trans-10-undecenoic acid methyl ester 23 trans-10-undecenoic acidethyl ester 36 n-octanoic acid 37 n-octanoic acid methyl ester 38n-octanoic acid ethyl ester 47 trans-2-decenoic acid- trans-2-decenylester 48 trans-2-decenoic acid cyclohexyl ester 53 trans-2-decenoic acidoctyl ester 54 trans-2-decenoic acid isopropyl ester 62 trans-3-decenoicacid methyl ester 63 trans-3-decenoic acid ethyl ester 77 geranic acidethyl ester 78 3,7-dimethyloctanoic acid ethyl ester

From FIGS. 1 to 3, the compounds shown in Table 1 showed numericalvalues of one or more times (except for compound 21), and MAP kinaseswere activated (phosphorylated). In particular, trans-2-decenoic acidesters such as any of the compounds having different alkoxy groups(compound Nos. 7, 8, 47, 48, 53 and 54) showed twice or more numericalvalues, and high MAP kinase activation (phosphorylation) effects wereshown. In particular, trans-2-decenic acid ethyl ester was as high as 11times, and secondarily, trans-2-decenic acid methyl ester was 6 times.Also, decanoic acid methyl ester, decanoic acid ethyl ester,trans-2-decenoic acid, trans-9-decenoic acid methyl ester,trans-9-decenoic acid ethyl ester, trans-10-undecenoic acid methylester, trans-10-undecenoic acid ethyl ester, geranic acid ethyl esterand 3,7-dimethyloctanoic acid ethyl ester showed high activities.

Example 2 Behavior Test of Mice Under Stress (1) Stress

Mice under established chronic mild stress (hereinafter referred to as“stress”) were used for a depression and anxiety disorder-like modelanimal. Stress was applied as follows. 7-week-old male ddY strain micefed in one cage for 4 mice were used and first subjected to forcedswimming for 15 minutes. Then, the mice were fed in a sloped cage for 2days, and then rested in a normal cage for 1 day. Then, the mice werefed in a dirt cage moisturized with 200 mL of water for 1 day and thenrested in a normal cage for 1 day. Then, the mice were fed in arotational cage that rotated at 180 rpm for 1 day, and then rested in anormal cage for 1 day. Stress taking for 1 week using these 3 kinds ofcages was regarded as 1 cycle, and the stress was applied for 3 cycles(3 weeks). At the same time as stress, the compound 8 that was dissolvedin 0.1% DMSO to adjust a concentration so as to have each of the dosesof 0, 20, 100 and 500 μg/kg was intraperitoneally injected for 3 weeksin an amount of 0.25 mL in each time once per day. The compound 8 wasalso administered to non-stressed examples for 3 weeks in the samemanner. For 0 μg/kg administration examples of controls, a phosphoricacid buffer containing 0.1% DMSO (controls were expressed as “PBS” or “0μg/kg” in the figure shown below) was administered in an amount of 0.25mL to each of a stressed example and a non-stressed example. Thebehavior test of mice was performed on each of the following: anadministration group to which the compound 8 was administered at thesame time as stress for 3 weeks and an administration group to which thecompound 8 was administered for 3 weeks without stress (hereinafterreferred to as the “stressed 3 week-administration groups”); and anadministration group to which the compound 8 was administered at thesame time as stress for 1 week and an administration group to which thecompound 8 was administered for 1 week without stress (hereinafterreferred to as the “stressed 1 week-administration groups”).Additionally, an example in which stress was applied to a mouse may bereferred to as a stressed example, and an example in which stress wasnot applied to a mouse maybe referred to as a non-stressed example.Further, a significance test was performed on the results of Examples 1to 7 in a Student's t-test.

(2) Behavior Test 1. Burying the Glass-Marbles Test

Tips were bedded so as to have a height of 5 cm in a box with 30 m² and25 glass marbles (1.6 cm in diameter) were placed thereon at 5cm-interval and behaviors of mice were observed. The number of buriedglass marbles was found by counting each glass marble that was buried in⅔ or more from visual observation from just above after 15 minutes asone count. The test was performed for 3 hours. Since a mouse understress having an anxiety symptom (condition) takes such a behavior asburying glass marbles, a level of the anxiety symptom can be evaluatedfrom the number of buried glass marbles.

FIGS. 4 and 5 show the results.

The numbers of glass marbles buried by mice had a significant differencebetween a stressed example and a non-stressed example of controls (PBS)in any of “the stressed 3 week-administration group” and “the stressed 1week-administration group”; on the contrary, there was no significantdifference between a stressed example and a non-stressed example in eachadministration example of the administration groups of the compound 8(see FIGS. 4 and 5), and the compound 8 suppressed an anxiety symptom.From the result of “the stressed 1 week-administration group”, thecompound 8 is considered to immediately act on the anxiety symptom.

2. Tail Suspension Test

A site of 1 cm from the tip of the tail of a mouse was grasped by ahand, being apart from a floor from 10 cm, and an immobility time insuspension for 6 minutes was measured for 2 hours. Since the immobilitytime prolongs when the mouse has a depressive symptom (condition), alevel of the depressive symptom (condition) can be evaluated from thelength of the immobility time. FIGS. 6 and 7 show the results.

The immobility times of mice had a significant difference between astressed example and a non-stressed example of controls (PBS) in any of“the stressed 3 week-administration groups” and “the stressed 1week-administration groups”; on the contrary, there was no significantdifference between a stressed example and a non-stressed example in eachadministration example of the administration groups of the compound 8(see FIGS. 6 and 7), and the compound 8 suppressed a depressive symptom.From the result of “the stressed 1 week-administration group”, thecompound 8 is considered to immediately act on the depressive symptom.

3. Elevated Plus Maze Test

Using a device having an elevated maze cruciately crossed, in which 10cm-wall was provided in the X axis (closed arm, 30 cm×10 cm) and no wallwas provided in the Y axis (open arm, 30 cm×10 cm), being apart from 50cm from a floor, behaviors of mice in 5 minutes were observed to measurea time for being in the open arm and the number of times of entering theclosed arm. In this test, an anxiety level can be evaluated from thetime for being the open arm, and a behavior amount can be evaluated fromthe number of times of entering the closed arm. From the evaluations,when a mouse has an anxiety symptom (condition), the time for being inthe open arm prolongs, and thus, a level of the anxiety symptom(condition) can be evaluated from the length of the time. FIGS. 8 to 11show the results.

Since the numbers of times when mice entered the closed arm had nosignificant difference between the control (PBS) and the compound 8administration groups and between the stressed groups and thenon-stressed groups (see FIGS. 9 and 11), administration of the compound8 did not give an effect on behavior amounts of the mice, and thus, thetimes when the mice were in the open arm did not depend on the behavioramounts. The times when the mice were in the open arm had a significantdifference between stressed examples and non-stressed examples ofcontrols (PBS) in any of “the stressed 3 week-administration groups” and“the stressed 1 week-administration groups”, and there was nosignificant difference between stressed examples and non-stressedexamples of each administration example in the compound 8 administrationgroups (see FIGS. 8 and 10), and the compound 8 thus suppressed theanxiety symptom of mice. From the result of “the stressed 1week-administration group”, the compound 8 is considered to immediatelyact on the anxiety symptom.

Example 3 Measurement of Weights of Mice Before and After Stress

A weight of a mouse before stress was subtracted from the weight of themouse immediately after administering the compound 8 as well as stressfor 3 weeks to obtain a weight gap (g). FIG. 12 shows the result.

FIG. 12 reveals that a weight of a non-stressed example of the control(PBS) increased, and on the other hand, a weight of a stressed examplesignificantly decreased due to depressive and anxiety symptoms. On theother hand, ratios of the weight decreases in stressed examples of thecompound 8 administration groups were less due to a suppression effectof the compound 8 on depressive and anxiety symptoms as compared to thecontrol (PBS), and there was no significant difference between astressed example and a non-stressed example in the 500 μg/kgadministration group and an effect on the body weights was notrecognized.

Example 4 Measurement of Activation of MAP Kinase with Compound 8

After 24 hours from completion of stress for 3 weeks in the same methodas described in Example 2, mice administered with the compound 8 indoses of 0, 20, 100 and 500 μg/kg respectively at the same time asstress and non-stressed mice administered with only a phosphoric acidbuffer containing 0.1% DMSO were sacrificed and the cortexes of frontallobes and hippocampi were excised respectively, and thereto were addedRIPA buffers in 19 times of an amount of each wet weight.

Then, sonication was performed by an ultrasonic grinder until tissuescannot be seen, centrifuged at 12000 rpm for 15 minutes, and thesupernatant was recovered to form a tissue protein extraction solution.The protein concentration of the extraction solution was measured usinga BCA Protein Assay Kit (TAKARA BIO INC.), each of extraction solutionscontaining 3 μg for a MAP kinase and 5 μg for a phosphorylated MAPkinase was added with a ⅓ amount of a sample buffer for electrophoresisat 4 times of a concentration, and a 1/10 amount of 2-mercaptoethanoland thermally treated at 95° C. for 5 minutes and subjected toelectrophoresis with a 10% SDS polyacrylic amide gel. Proteins were thentranscribed to a PVDF membrane from the gel and the membrane aftertranscription was immersed in a block liquid (TBS containing 5% skimmedmilk) for 1 hour, and reacted with primary antibodies: an anti-MAPkinase antibody (Cell Signaling Technology, Inc.) and anantiphosphorylated MAP kinase antibody (Cell Signaling Technology, Inc.)overnight, and further reacted with the secondary antibody: an alkalinephosphatase labeled anti-rabbit IgG antibody (Promega KK) for 1 hour.Lastly, a D1G3 buffer was used as the substrate of an alkalinephosphatase and incubated with a liquid obtained by adding nitrobluetetrazolium (NBT) and 5-bromo-4-chloro-3-indolylphosphate p-toluidinesalt (BCIP) for several minutes to develop a color. For a concentrationmeasurement of a band, an intensity was calculated with Image J (BioArtsInternational, Inc.) to find a phosphorylated MAP kinase amount of eachof the compound 8 administration groups to a phosphorylated MAP kinaseamount in a non-stressed example. FIGS. 13 and 14 show the results.

FIGS. 13 and 14 reveal that the phosphorylated MAP kinase amount of thecontrol (0 μg/kg) significantly decreased in both of the cortex offrontal lobe and hippocampus as compared to a non-stressed example. Onthe other hand, the compound 8 administration groups had anadministration example in which the phosphorylated MAP kinase amountsignificantly increased as compared to the control (0 μg/kg) and thecompound 8 promoted activation (phosphorylation) of the MAP kinase.

Example 5 mRNA Expression of Neurotrophic Factor and GlucocorticoidReceptor in Cortex of Frontal Lobe and Hippocampus of Mouse

mRNA expressions of a brain-derived neurotrophic factor (hereinafterexpressed as “BDNF”), neurotrophin-3 (hereinafter expressed as “NT-3”),a nervous growth factor (hereinafter expressed as “NGF”) and aglucocorticoid receptor (hereinafter expressed as “GR”) in a cortex offrontal lobe and a hippocampus of a mouse after stress were examined asfollows. After 24 hours from completion of stress for 3 weeks in thesame method as described in Example 2, mice administered with thecompound 8 in each of the amounts of 0, 20, 100 and 500 μg/kg, at thesame time as stress, and non-stressed mice administered with only aphosphoric acid buffer containing 0.1% DMSO were sacrificed and thecortexes of frontal lobes and hippocampi were excised respectively, andthereto were added TriZol. The both tissues were crashed with ahomogenizer respectively, chloroform was then added thereto, and themixture was centrifuged at 15000 rpm for 15 minutes to take out theaqueous phase. This aqueous phase was added with isopropanol andcentrifuged at 15000 rpm for 15 minutes.

The obtained precipitate was washed with 70% ethanol, dried and thendissolved in TE to form a total RNA sample. For reverse transcription ofRNA, annealing was performed by adding a CDS primer (oligo dt primer)solution to 0.5 μg of the total RNA sample to react at 72° C. for 2minutes. Then, a 5× first strand buffer, DTT, dNTP and a reversetranscriptase were added thereto to react at 42° C. for 60 minutes.Thereto was added TE and reacted at 72° C. for 7 minutes to form a cDNAsample. A PCR reaction was carried out by adding a mixed solutioncontaining cDNA, Taq polymerase and primers (respective primers havingsequences (10 to 20) specific to BDNF, NGF, NT-3 and GR). In the PCRreaction, a treatment was carried out at 94° C. for 5 minutes ininitiation, modification at 94° C. for 45 seconds, annealing at 61° C.for 45 seconds, and elongation at 72° C. for 30 seconds were performedas one cycle, 28 cycles were repeated for BDNF, 30 cycles were repeatedfor NT-3, 34 to 37 cycles were repeated for NGF, and 37 cycles wererepeated for GR. At last, an elongation reaction was performed at 72° C.for 7 minutes. After the PCR reaction, electrophoresis was carried outwith a 2% agarose gel and an amplified DNA fragment was subjected tofluorescent color development of a band with ethidium bromide. Thefluorescence intensity of the band was read with a densitometer todigitalize and the obtained value was divided with a fluorescenceintensity of a band of RT-PCR of β actin mRNA of an internal standard,which has been found separately, and a graph was formed assuming thatthe band value of a non-stressed mouse/β actin band value was 1.0. FIGS.15 and 16 show the results.

Expressions of mRNAs of BDNF, NT-3 and GR in the cortexes of frontallobes were significantly decreased in controls (0 μg/kg) as compared tonon-stressed examples (see FIG. 15), and Expressions of mRNAs of BDNF,NGF and GR in the hippocampi were significantly decreased in controls (0μg/kg) as compared to non-stressed examples (see FIG. 16). Since it hasalready been known that expressions of mRNAS of GR and BDNF are lowered,stress of the test provoking reduction in expression of mRNAs of BDNFand GR is determined to be homogeneous to conventionally reportedstress. In FIGS. 15 and 16, since mRNAs of BDNF, NT-3, NGF and GR werereturned to the level of non-stressed examples in the compound 8administration examples, reduction in expression of neurotrophic factorrelated genes accompanied by stress was suppressed with the compound 8,and considered to be improved to a state of a non-stressed example.

Example 6 Behavior Test of Mice After Completion of Stress

Example 2 is a behavior test in which the compound 8 was administered toa mouse as well as stress was applied and a suppression effect of thecompound 8 on depressive and anxiety symptoms of the mouse was examined.On the other hand, the present example is a behavior test in which onlystress was applied for 3 weeks without administering the compound 8 andstress was completed and a therapeutic effect of the compound 8 ondepressive and anxiety symptoms of a mouse after removing stress wasexamined. Stress was applied to a mouse in the same manner as in Example2. For behavior tests of mice, the burying the glass-marbles test, thetail suspension test and the elevated plus maze test were performedrespectively in the same manners as in Example 2 on groups withoutadministering the compound 8 immediately after completion of stress for3 weeks and groups without administering the compound 8 with no stressfrom the start (hereinafter referred to as “non-administration groupsafter completion of stress”); a group administered with the compound 8for 1 week after completion of stress and a group administered with thecompound 8 for 1 week in the same manner as the former group with nostress from the start (hereinafter referred to as “1 week-administrationgroups after completion of stress”); and a group administered with thecompound 8 for 2 weeks after completion of stress and a groupadministered with the compound 8 for 2 weeks in the same manner as theformer group with no stress from the start (hereinafter referred to as“2 week-administration groups after completion of stress”). In any ofthe following behavior tests of mice, there were significant differencesbetween a stressed example and a non-stressed example of controls (0μg/kg) of “the non-administration groups after completion of stress” andbetween a stressed example and a non-stressed example of the compound 8administration groups (see FIGS. 17, 20 and 23). Although doses aredescribed in FIGS. 17, 20, 23 and 24 showing “the non-administrationgroups after completion of stress”, the compound 8 was not administeredin any of the groups as described above, and mice corresponding to eachof the administration groups, “the 1 week-administration groups aftercompletion of stress” and “the 2 week-administration groups aftercompletion of stress” are only shown.

1. Burying the Glass-Marbles Test

The numbers of glass marbles buried by mice had a significant differencebetween a stressed example and a non-stressed example of controls (0μg/kg) in any of “the 1 week-administration groups after completion ofstress” and “the 2 week-administration groups after completion ofstress”; on the other hand, there was no significant difference betweena stressed example and a non-stressed example in each administrationexample of 20 μg/kg and 100 μg/kg (see FIGS. 18 and 19), and thecompound 8 exhibited a therapeutic effect on an anxiety symptom afterstress was removed. The result of “the 1 week-administration groupsafter completion of stress”, the compound 8 is considered to have animmediate therapeutic effect on the anxiety symptom.

2. Tail Suspension Test

The immobility times of mice had a significant difference between astressed example and a non-stressed example of controls (0 μg/kg) in“the 1 week-administration groups after completion of stress”, but therewas no significant difference between a stressed example and anon-stressed example in 500 μg/kg administration examples (see FIG. 21),and the compound 8 exhibited an immediate therapeutic effect on ananxiety symptom after removing stress. The immobility times of mice hada significant difference between a stressed example and a non-stressedexample of controls (0 μg/kg) in “the 2 week-administration groups aftercompletion of stress”, but there was no significant difference between astressed example and a non-stressed example in all administrationexamples (see FIG. 22), and the compound 8 exhibited a high therapeuticeffect on a depressive symptom after removing stress.

3. Elevated Plus Maze Test

Since the numbers of times when mice enter the closed arm had nosignificant difference between controls (0 μg/kg) and the compound 8administration groups and between stressed groups and non-stressedgroups in any of “the non-administration group after completion ofstress”, “the 1 week-administration groups after completion of stress”and “the 2 week-administration groups after completion of stress” (seeFIGS. 24, 26 and 28), administration of the compound 8 did not give aneffect on behavior amounts of the mice, and thus, the times when themice are in the open arm do not depend on the behavior amounts. Thetimes when the mice were in the open arm had a significant differencebetween a stressed example and a non-stressed example of the controls (0μg/kg) in “the 1 week-administration groups after completion of stress”,but there was no significant difference between a stressed example and anon-stressed example of the 500 μg/kg administration examples (see FIG.25), and the compound 8 exhibited an immediate therapeutic effect on ananxiety symptom after stress was removed. There was a significantdifference between a stressed example and a non-stressed example of thecontrols (0 μg/kg) in “the 2 week-administration group after completionof stress”, but there was no significant difference between stressedexamples and non-stressed examples of all administration examples (seeFIG. 27), and the compound 8 exhibited a high therapeutic effect on ananxiety symptom after stress was removed.

Example 7 Measurement of Weights of Mice After Completion of Stress

A weight of a mouse before stress was subtracted from of the weight ofthe mouse immediately after completion of stress for 3 weeks withoutadministering the compound 8 and the weight of the mouse afteradministering the compound 8 for 1 week after completion of stress,respectively, to obtain weight gaps (g). FIGS. 29 and 30 show theresults. Although doses are described in FIG. 29, the compound 8 was notadministered as described above, and mice corresponding to each of theadministration groups administered with the compound 8 for 1 week aftercompletion of stress are only shown.

From FIG. 29, the weight gaps of mice immediately after completion ofstress had significant differences between a stressed example and anon-stressed example of the controls (0 μg/kg) and between stressedexamples and non-stressed examples corresponding to respectiveadministration groups, but in the 500 μg/kg administration examplesafter administrating the compound 8 for 1 week after completion ofstress, there was no significant difference between a stressed exampleand a non-stressed example due to a therapeutic effect of the compound 8on repressive and anxiety symptoms.

Example 8 Comparison of Effects of Compound 8 and Fluvoxamine onDepressive Symptom in Tail Suspension Test

7-week-old male ddY strain mice fed in one cage for 4 mice were firstsubjected to forced swimming for 15 minutes in the same manner as stressin Example 2 (behavior test of mice under stress) (1) described in theparagraph [0063] in the specification. Then, stress for 1 week using 3kinds of cages was regarded as 1 cycle, and the stress was applied for 3cycles (3 weeks) and chronic mild stress (CMS) was applied. The compound8 and fluvoxamine were administered to mice immediately after stress inExp. 1, 1 week after stress in Exp. 2, and 2 weeks after stress in Exp.3, respectively, and tail suspension tests were carried out on therespective test groups. In each test group, the compound 8 was adjustedin concentration so as to have each of the doses of 20, 100 and 500μg/kg and fluvoxamine was adjusted in concentration so as to have a doseof 1000 μg/kg, by dissolving in 0.1% DMSO, and each dose of the compound8 and fluvoxamine were intraperitoneally injected to mice once per dayfrom the time immediately after stress. Further, to 7-week-old male ddYstrain mice fed in one cage for 4 mice without stress, the compound 8and fluvoxamine were administered to carry out the tail suspension testsin the same manner as the stressed mice. FIG. 31 show the result. Asignificance test was performed in a Student's t-test (*). FIG. 31Ashows an evaluation method of the behavior test (behavior assessment).FIG. 31B shows immobility times of the mice. “DAEE” denotes the compound8 and “Flu” denotes fluvoxamine.

FIG. 31B reveals that the compound 8 had an effect in a dose of 500μg/kg in Exp. 2 (1 week after stress) but an effect could not berecognized with fluvoxamine. An effect of fluvoxamine was recognizedonly in Exp. 3 (2 weeks after stress). The results found that thecompound 8 was excellent in an effect on a depressive symptom.

Example 9 Voluntary Alternation Behavior Test of Mice Administered withTrimethyltin (Y-Maze Test)

The voluntary alternation behavior test (Y-maze test) is one kind of abehavior test using a Y-shaped maze constituted with three arms. First,a mouse is placed on the tip of one arm out of the three arms, lettingthe mouse freely explore in the maze over 8 minutes, and arms where themouse transferred are recorded in order. Then, the number of times oftransferring to each arm (total arm selection number (total armentries)) and the number of sequentially selecting three different arms(number of alternation behavior (number of spontaneous behavior)) arecounted. A normal mouse shows a high probability that indicates abehavior of transfer with selecting three different arms, and a mousewith declined memory and learning abilities shows a low probability oftaking such a behavior, and thus, number of spontaneous behavior÷(totalarm entries−2)×100=alternation behavior (%) was calculated to be theindex of the voluntary alternation behavior.

Trimethyltin (hereinafter referred to as “TMT”) is an organic metalcompound having neurotoxicity, and acute nerve cell death is casedparticularly in granular nerve cells of the hippocampus dentate gyrus,which thus affects memory and learning abilities of a mouse. Therefore,TMT is administered to a mouse, and the alternation behavior (%) issignificantly decreased.

The voluntary alternation tests (Y-maze tests) were performed on 1) testgroups to which the compound 8 was singly administered and 2) testgroups to which TMT and the compound 8 were administered. In the testgroups to which the compound 8 was singly administered, the compound 8was dissolved in 0.1% DMSO to adjust a concentration so as to have eachof the doses of 100 and 500 μg/kg and intraperitoneally injected to 6mice in every administration example each in a dose of 0.25 mL. In 0μg/kg weight administration examples of controls, 0.25 mL of aphosphoric acid buffer containing 0.1% DMSO was intraperitoneallyinjected in each time. The voluntary alternation tests (Y-maze tests)were performed on each of the administration groups after the elapse of2 hours from intraperitoneal administration. In the test group in whichTMT and the compound 8 were administered, TMT (2 mg/kg weight) wasintraperitoneally administered to a mouse, and the compound 8 wasintraperitoneally administered in the same manner as the test groups inwhich the compound 8 was singly administered after 2 days have passed,and the voluntary alternation test (Y-maze test) was performed after theelapse of 2 hours. FIG. 32 show the results. Significance tests wereperformed in a Turkey's test (*, ##) and a Student's t-test (++). “DAEE”denotes the compound 8.

According to FIG. 32B, the total arm entries had no difference amongrespective test groups, which means that there was no difference inmotor functions of the respective test groups. FIG. 32A reveals that, inthe test examples in which TMT and the compound 8 were administered atthe same time, administration of the compound 8 suppressed decrease inthe voluntary alternation behaviors that significantly decreased due toTMT treatments. On the other hand, it was found that, also in the testexamples in which the compound 8 was singly administered, the voluntaryalternation behaviors significantly increased due to administration ofthe compound 8, and thus, the compound 8 can impart resistivity againstneurotoxicity of TMT to suppress declined memory and learning abilitiesand to promote the memory and leaning abilities of mice. From the aboverespects, the compound 8 is considered to have an activity of improvingmemory and learning abilities by further increasing general functions ofthe hippocampus. Such an activity is considered to give a suppressiveeffect also on declined nervous functions due to an amyloid β peptide inAlzheimer's disease.

Example 10 Novel Object Recognition Test on Mice

A promotion activity of brain functions due to the compound 8 wasexamined using a novel object recognition test. The novel objectrecognition test is a method of evaluating memory and learning abilitiesfrom behaviors after showing a “familiar object” and a “novel object” tomice, utilizing the fact that the mice take exploration behaviors(behaviors such as coming closer to or smelling an object) when theyfind the “novel object”.

The compound 8 was dissolved in 0.1% DMSO to adjust a concentration soas to have each dose of 100 and 500 μg/kg and intraperitoneally injectedto 6 mice in every administration example each in a dose of 0.25 mL. Ina 0 μg/kg weight administration example of a control, 0.25 mL of aphosphoric acid buffer containing 0.1% DMSO was intraperitoneallyinjected. The compound 8 was administered to mice and the novel objectrecognition test was performed after 2 hours. That is, a mouse was putin a place where two identical objects were present for 15 minutes tolet the mouse memorize the two objects (these two objects will be“familiar objects”). After the elapse of 1 day, an exploration behaviortest in which the mouse was put in a place where the “familiar objectsand “novel objects” were present was carried out for 10 minutes. FIG. 33show the results. Significance tests were performed in a Student'st-test (*) and a Turkey's test (##). “DAEE” denotes the compound 8.

FIG. 33A shows time for the mice to explore “familiar objects” and“novel objects” (exploration time). FIG. 33B shows recognition indices.The recognition indices are expressed by Y/X+Y assuming that a time forexploring the “familiar objects” is X and a time for exploring the“novel objects” is Y. It was found from FIG. 33B that since therecognition indices of the “novel objects” became high in examples towhich the compound 8 was administered, the compound 8 had an activity ofenhancing memory and learning abilities of a healthy animal. Such anactivity is considered to give a suppressive effect also on declinednervous functions due to an amyloid β peptide in Alzheimer's disease.

1. An agent having a neurotrophic factor-like activity, comprising, asan active ingredient, any one compound included in fatty acids eachhaving 8 carbon atoms (C8) or having 10 carbon atoms (C10) to 12 carbonatoms (C12) or fatty acid esters thereof, such as n-octanoic acid methylester, n-octanoic acid ethyl ester, 3,7-dimethyloctanoic acid ethylester, or geranic acid ethyl ester, each of which has 8 carbon atoms(C8), decanoic acid methyl ester, decanoic acid ethyl ester,trans-2-decenoic acid, trans-2-decenoic acid methyl ester,trans-2-decenoic acid ethyl ester, trans-2-decenoic acid-2-decenylester, trans-2-decenoic acid cyclohexyl ester, trans-2-decenoic acidoctyl ester, trans-2-decenoic acid isopropyl ester, trans-3-decenoicacid methyl ester, trans-3-decenoic acid ethyl ester, trans-9-decenoicacid, trans-9-decenoic acid methyl ester, or trans-9-decenoic acid ethylester, each of which has 10 carbon atoms (C10), trans-10-undecenoic acidmethyl ester or trans-10-undecenoic acid ethyl ester, each of which has11 carbon atoms (C11), and dodecanoic acid, dodecanoic acid methylester, or dodecanoic acid ethyl ester, each of which has 12 carbon atoms(C12), or salts thereof or prodrugs thereof.
 2. The agent having aneurotrophic factor-like activity according to claim 1, which is apreventive and therapeutic agent for a nervous disorder.
 3. The agenthaving a neurotrophic factor-like activity according to claim 2, whereinthe nervous disorder is a neurodegenerative disease.
 4. The agent havinga neurotrophic factor-like activity according to claim 3, wherein theneurodegenerative disease is Alzheimer's disease, Parkinson's disease,amyotrophic lateral sclerosis (ALS), Huntington's disease, progressivesupranuclear palsy (PSP), or diabetic neuropathy.
 5. The agent having aneurotrophic factor-like activity according to claim 2, wherein thenervous disorder is a mental disease.
 6. The agent having a neurotrophicfactor-like activity according to claim 5, wherein the mental disease isdepression.
 7. The agent having a neurotrophic factor-like activityaccording to claim 5, wherein the mental disease is anxiety disorder(neurosis).
 8. Trans-2-decenoic acid-2-decenyl ester, having aneurotrophic factor-like activity.
 9. Trans-2-decenoic acid cyclohexylester, having a neurotrophic factor-like activity.